What guarantees are associated with your TriFECTa kit? Is the designed RNA duplex specific?
IDT guarantees that at least two of the three Dicer-Substrate duplexes in the splice-common TriFECTa® kit will give a least 70% knockdown of the target mRNA when used at 10 nM concentration and assayed by quantitative RT-PCR when the fluorescent transfection control duplex indicates that >90% of the cells have been transfected and the HPRT positive control works with the expected efficiency. Our Bioinformatics department did a full screening of the recommended sequences before adding them to the database. This included BLASTing the sequences to ensure they only line up with the gene in question, performing secondary structure checks, and making sure the end stabilities were favorable for incorporation into the RISC complex.